Diabetic Uropathy is a term for a range of debilitating urologic complications such as bladder dysfunction, urinary incontinence, urinary tract infection and sexual dysfunction, that are among the most common and costly, yet understudied complications of diabetes mellitus (DM), an incurable disease that affects at least 20 million people in the U.S. and is rising in prevalence with the rapidly rising prevalence of obesity. Therapeutic options for diabetic uropathy are inadequate and have not improved over the last 50 years. In response to RFA-DK-05-011, we propose to work with the Animal Models of Diabetic Complications Consortium's organizational structure to function as the 'Diabetic Uropathy Pathobiology Site" to participate in development of two novel mice models of diabetic uropathy and to investigate the mechanisms of the pathophysiology of diabetic bladder dysfunction in these animals. Based on the observed temporal effects of diabetes on the bladder function in small animals, we hypothesize that depletion of manganese superoxide dismutase (MnSOD) specifically in smooth muscle of adult mice will exacerbate accumulation of free radicals in smooth muscle during STZ-induced diabetes and accelerate the onset of the decompensated phase of diabetic bladder dysfunction. We hypothesize further that limiting depletion of MnSOD to arterial smooth muscle will have a lesser effect on STZ-induced diabetic bladder dysfunction by limiting exacerbationof STZ-induced free radical accumulation to the vasculature. Depletion of MnSOD selectively in total and arterial smooth muscle in the MnSODlox/lox, SM- CreER12 mice and MnSODlox/lox, ASM-CreERT2 mice, respectively, will be accomplished by administration of 4-hydroxytamoxifento activate Cre recombinase expressed in the smooth muscle. The animals will be further treated with 4-hydroxytamoxifen treatment, and half of them will be injected with STZ to induce diabetes. The bladder function in the animals will be studied via four specific aims to examine: 1) the temporal alterations in the in-vivo bladder function by micturition habits and conscious cystometry;2) the temporal course of morphological changes in diabetes-induced bladder hypertrophy;3) temporal alterations in the contractile function of the detrusor muscle;4) the temporal alterations in afferent and efferent autonomic pathways innervating the bladder. The Principal Investigator and the research team have a productive track record in being a part of the existing AMDCC since 2003 and have functioned well under the auspices of the NIH, and the Steering and External Advisory Committees of the AMDCC. PERFORMANCE SITE(S) (organization, city, state) Cleveland Clinic Lerner College of Medicine of CWRU (CCLCM) 9500 Euclid Avenue Cleveland, Oh 44195 PHS 398 (Rev. 09/04) Page 2 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): Feldman, Eva L. KEY PERSONNEL. See instructions. Use continuation pages as neededto provide the required information Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Daneshgari, Firouz FirouzDaneshgari CCLCM Liu, Guiming NA CCLCM Powell, C. Thomas NA CCLCM Ukimura, Osamu NA CCLCM in the format shown below. Role on Project Principal Investigator Research associate Research Associate Research Scholar OTHER SIGNIFICANT CONTRIBUTORS Name Bushman, Wade Brown, Jeanette Bhat, Manjue Charron, Maureen Katims, Jefferson Organization Role on Project University of Wisconsin Consultant UCSF Consultant CCLCM Consultant Albert Einstein College of Med Consultant Neurotron, Inc. Consultant Human Embryonic Stem Cells [X]No d Yes If the proposed project Involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://stemcells.nih.qov/reqistrv/index.asp. Usecontinuation pages as needed. If a specific line cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line Disclosure Permission Statement. Applicable to SBIR/STTR Only. See SBIR/STTR instructions. Yes No PHS 398 (Rev. 09/04) Page U Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Principal Investigator/Program Director (Last, First, Middle): Daneshgari, FifOUZ The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page . 1 Description, Performance Sites, Key Personnel, Other Significant Contributors, and Human Embryonic Stem Cells; Table of Contents Detailed Budget for Initial Budget Period (or Modular Budget) 4-5 Budget for Entire Proposed Period of Support (not applicablewith Modular Budget) Budgets Pertaining to Consortium/Contractual Arrangements (not applicablewith ModularBudget) Biographical Sketch - Principal Investigator/ProgramDirector (Not to exceed four pages) 6-8 Other Biographical Sketches (Not to exceed four pages for each - Seeinstructions) 9-32 Resources 33 Research Plan, 34-58 Introduction to Revised Application (Not to exceed 3pages;SBIR/STTR Phase I notto exceed 1page.).. Introduction to Supplemental Application (Not to exceed one page) A. Statement of Intent 34_ B. General Goals and Background 34_ C. New Mouse Models ^. (Items A-D: not to exceed 25pages*) 35-36 Phase I Progress Report (SBIR/STTR Phase II ONLY) f * SBIR/STTR Phase I: Items A-D limited to 15pages. D. Pathophysiology and Organ-Specific Phenotyping 37-58 E. Human Subjects Research 59 Protection of Human Subjects (Required if Item 4 on the Face Page is marked "Yes") Data and Safety Monitoring Plan (Required if Item 4 on the Face Page is marked "Yes" and a Phase I, II, or III clinical trial is proposed) '. Inclusion of Women and Minorities (Required if Item 4 on the Face Page is marked "Yes" and is Clinical Research) Targeted/Planned Enrollment Table (for new and continuing clinical research studies) Inclusion of Children (Required if Item 4 on the Face Page is marked "Yes") F. Vertebrate Animals 59-60 G. Literature Cited 61-65 H. Consortium/Contractual Arrangements 66_ I. Resource Sharing .-.. '.;66_ J. Letters of Support (e.g., Consultants) 67-74 Commercialization Plan (SBIR/STTR Phase II and Fast-Track ONLY) Checklist, 75_ Check if Appendix (Five collated sets. No page numbering necessary for Appendix.) Appendix is Included Appendices NOT PERMITTED for Phase I SBIR/STTR unless specifically solicited.. Number of publications and manuscripts accepted for publication'(not to exceed 10) Other items (list): PHS 398 (Rev. 09/04) Page 3 Form Page 3